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Valorar el comportamiento histológico, en un modelo animal de conejo, de un nuevo cemento óseo modificado, el cual aumenta la liberación local de antibiótico, en la infección ósea.

The aim of this study was to evaluate the in vitro response of mesothelial cells (MC) in terms of their ability to cover different biomaterials. MC were harvested from human omentum. The MC from the first passage were seeded onto different biomaterials from 10 min to 24 h: PL-PU99 (polypropylene-polyurethane); DM (ePTFE); PL (polypropylene); and PL + Col (polypropylene-collagen). The prosthetic surface covered was examined by microscopy and quantified. PL-PU99: The MC were adhered to the biomaterial 10 min post-incubation. At 4 h, the 53.12±7.86% of the prosthesis were coated with polygonal cells. At 12 h, 96.32±11.32% of the biomaterial was coated. DM: between 30 min to 8 h, the MC cells form small, round colonies. At 12 h, polygonal and fusiform secretory cells were observed (68.94±5.78%). 93.54±11.49% of surface was coated after 24 h. PL: only isolated cells were observed on the prosthesis. PL + Col: MC form a monolayer over prosthetic surface after 18 h (90.21±9.76). We conclude: (a) MC formed a stable monolayer over all the biomaterials tested with the exception of the PL due to its porosity. (b) The PL-PU99 showed the greatest potential for in vitro mesothelialization compared to the PL-Col and DM prostheses.

Objective:to evaluate the condition of organ donor arteries subjected to prolonged cold-ischaemia followed by cryopreservation, for their possible use as vascular grafts.

This study was designed to test a slow, controlled, automated process for the thawing of cryopreserved arteries, whereby specimen warming is synchronized with the warming of its environment. Segments of minipig iliac artery, 4–5 cm in length, were subjected to controlled, automated cryopreservation in a biological freezer at a cooling rate of 1°C/min to −120°C, followed by storage in liquid nitrogen at −196°C for 30 days. Following storage, the arterial segments were subjected to rapid (warming rate of approximately 100°C/min) or gradual (1°C/min) thawing. Thawed specimens were processed for light microscopy and for scanning and transmission electron microscopy, Cell death was determined by the TUNEL method. Metalloproteinase (MMP) expression was estimated by immunohistochemical analysis. Most of the cryopreserved vessels subjected to rapid thawing showed spontaneous fractures, mainly microfractures, whereas these were absent in slowly thawed specimens. In rapidly thawed vessels, the proportion of damaged cells was double that observed in those thawed more gradually. Increased intensity and extent of MMP-2 expression was shown by rapidly thawed specimens. The slow-thawing protocol tested avoids the formation of spontaneous fractures and microfractures and the accumulation of fluid within the arterial wall tissue. This results in improved tissue preservation.

Objective to extend present knowledge of the biomechanical and structural changes which occur in the cryopreserved, rapidly thawed arterial wall. Materials and methods minipig iliac arterial segments were cryopreserved at −196 °C in either minimum essential medium or Wisconsin solution. Fresh segments served as the control group. After 1 month, the specimens were rapidly thawed (37 °C) and processed for biomechanical, ultrastructural, morphological and immunohistochemical (MMP-1, MMP-2, MMP-3 and MMP-9) analysis. Visualisation of apoptotic cells was performed by TUNEL method. For the mechanical distension analysis, an in vitro circuit was designed. Results the cryopreserved segments showed a 42% incidence of spontaneous fracture and the appearance of microfractures which affected the endoluminal third of the vessel. An accumulation of liquid in the subelastica was observed. An increased expression of wall-degradative enzymes (mainly MMP-2) was also observed following cryopreservation. No significant differences were detected in the proportional elasticity module or tensile strength of the specimen groups. No differences in mechanical distension were observed between groups after the vessel segments were subjected to the pulsatile circuit flow for 72 h. Cell damage was most intense in the specimens cryopreserved in Wisconsin solution. Conclusions cryopreservation in both the solutions employed, followed by rapid thawing, induce changes in the permeability which increase the fragility of the cryopreserved arterial wall. Both increased expression of wall-degradative enzymes and accumulation of liquid may contribute to graft failure after implantation. Copyright 2000 Harcourt Publishers Ltd

Objective: An evaluation of the proteoglycan perlecan, collagen I and III, and metalloproteinases MMP−1, −2, −3 and −9 was performed to explore the possible relationships between ageing, affected vein region and reactive state of the varicose vein wall.Methods: Segments of saphenous vein were obtained from healthy subjects and from those with varicose veins. The vein specimens were subdivided according to subject age (<50/^50) and vein source (distal/proximal).Results: The walls of control vein specimens acquired a more collagenous appearance with age. These changes were not accompanied by significant modifications in the immunohistochemical markers used. In specimens from young patients, proximal varicose vein segments showed an increase in MMP-1, MMP-2 and MMP-9 expression. Subjects of more advanced age showed an increase in perlecan expression.Conclusion: This increase in MMPs could lead to the acceleration of the final fibrosclerotic process characteristic of the varicose vein wall.

Objectives:the aim of the present study was to investigate the changes which occur to the arterial wall following cryopreservation and thawing and to determine whether these changes are reversible after a week of culture in an organ bath.Materials and methods:rat iliac arterial segments were cryopreserved. Once thawed, the arterial segments were cultured for a period of 0, 1, 2, 4 or 7 days. Freshly isolated rat iliac vessels cultured for 7 days served as the control group. Evaluation was made of ultrastructural changes, the expression of metalloproteinase activity (MMP-1, MMP-3 and MMP-9) and the apoptotic state of cells.Results:the freezing-thawing process induced damage to the arterial segments compared to fresh control vessels. After 1 week of culture, arteries showed a high degree of tissue degeneration. Only a few individual endothelial cells remained on the luminal surface. There was a gradual increase in the proportion of apoptotic cells. The sequential expression of MMP-1 during the first 2 days and subsequent expression of MMP-3 and MMP-9 were of most significance.Conclusions:cryopreservation induced damage to the vessels which could not be reversed by organ culture. The changes observed in the expression of metalloproteinases may be indicative of the degenerative process which occurs in the extracellular matrix.

The objective of this work was to study the healing process at the interface between biomaterial and visceral peritoneum. Implants of polytetrafluoroethylene (ePTFE) and polypropylene prostheses were introduced into the abdominal wall of New Zealand rabbits. The behaviour of the biomaterials was analysed using light and scanning electron microscopy and immunohistochemistry in which a specific anti-rabbit macrophage monoclonal antibody (RAM 11) was employed. According to macroscopic observation, there was significantly fewer adhesions prosthesis-viscera to ePTFE than to polypropylene implants. After ePTFE implantation, restoration of the peritoneum took place in an orderly fashion. When polypropylene was used, the peritoneum formed was a disorderly tissue in which small areas of haemorrhage and necrosis could be seen to coincide with the appearance of adhesions. The number of labelled macrophages peaked 14 days after ePTFE or polypropylene implantation, after which it decreased gradually. It is concluded that, given the low rate of adhesion provoked by PTFE, this material is ideal for implants contiguous to the peritoneal cavity viscera. The macrophage response does not determine the use of one material or the other. The structure of the newly formed peritoneum and development of adhesions depends on the porosity of the biomaterial.

The postimplantation peritoneal adhesions formed with some biomaterials continue to represent a serious problem when these biomaterials are placed in contact with the organs of the abdominal cavity. The objective was to test the effects of phosphatidylcholine (PC) in the attempt to modulate the adhesive process, thus improving the biomaterial/visceral peritoneum interface. We have carried out an experimental study using the New Zealand white rabbit, implanting a 7 × 5 cm2polypropylene prosthesis into the anterior abdominal wall in such a way that it replaced all the tissue layers and was in contact with the intestinal loops. Three study groups were established: group I (control), in which only implantation of the biomaterial was performed; group II, in which the prosthesis was pretreated with a solution of PC; and group III, in which the implants were treated as in group II and the animals underwent intraperitoneal administration of 10ml of the same solution. The animals were killed on postimplantation day 14. The total surface of the prosthesis occupied by adhesions was measured and light and scanning electron microscopy were performed to analyse the healing process. In comparison with the controls, groups II and III did not present significant differences with respect to the resistance or extension of the adhesions. The microscopic results showed a rapid formation of disorderly and well-vascularised scar tissue enveloping the entire prosthesis. Thus, PC was unable to modulate the process of adhesion formation between the prosthesis and the organs of the abdominal cavity; nor did it induce changes in the cells of sufficient importance to alter the results of the healing process in the presence of this biomaterial.

Summary The authors have studied the histological, histochemical, and ultrastructural aspects of eight corticectomies of epileptogenic foci in the medial surfaces of frontal lobes.

Liposomes may exert certain toxic effects on human cells depending on their composition and concentration when used as drug carriers. In this study, the hehaviour of human fibroblasts in the presence of soya phosphatidylcholine (soya-PC) liposomes and the effect of the incorporation of vitamin E into the membrane of such liposomes, was investigated. Human fibroblasts were incubated for 72h with low (50μM) and high (200 μM) concentrations of large, unilamellar (LU V) soya-PC liposomes in the presence and absence of vitamin E (soya-PC:vitamin E; 10:0.5). The resulting cultures were studied by scanning and transmission electron microscopy, fluorescence microscopy and labelling with rhodamine B. The large unilamellar soya-PC liposomes obtained were of mean diameter 86.4nm (soya-PC) and 78.3 nm (soya-PC: vitamin E). No significant changes in cell viability were observed between untreated fibroblasts and those incubated with 50μm soya-PC liposomes. In contrast, a marked loss of cells was apparent at a concentration of 200 μM soya-PC liposomes which was reversible by the addition of vitamin E. Morphological changes including the production of microvilli or 'bubbling' of the cell membrane, were incurred by some of the fibroblasts on addition of soya-PC liposomes. The addition of vitamin E did not affect the changes produced at the lower liposome concentrations but was able to reduce the cytotoxic effects occurring at higher concentrations, possibly by inhibiting the formation of liposome aggregates. The use of PC-soya liposornes containing vitamin E at high concentrations is proposed as a means of delivering high doses of drugs.

AbstractThe use of liposomes as carriers for certain drugs minimizes the adverse effects of high drug concentrations. However, the liposomes themselves may exert toxic effects on human cells, depending on their composition and concentration. The aim of the present investigation was to compare the effects of dimyristoylphosphatidylcholine (DM-PC) liposomes with those of DM-PC liposomes containing α-tocopherol on human fibroblasts in culture. DM-PC liposomes had a dose-dependent effect on the growth of fibroblasts in culture, showing high toxicity at a phospholipid concentration of 300 μM. The incorporation of α-tocopherol in the liposome bilayer significantly improved cell survival in the presence of high DM-PC liposome levels. The results demonstrate that a-tocopherol is able to reduce the negative effects of high phospholipid concentrations and perhaps permit the use of liposome concentrations that would otherwise be toxic.

In this study we compared the behaviour of the non-porous on one side ePTFE Dual Mesh® prosthesis and the macroporous polypropylene mesh Marlex® in the repair of abdominal wall defects in rabbits. We evaluated the degree of integration with recipient tissue, biological tolerance, adhesion formation with viscera and the biomechanical resistance of the repair zone. Our results showed good biological tolerance of both prostheses and a high degree of adhesion formation in Marlex implants. In animals with Dual Mesh(r) implants, only loose adhesions were seen. Marlex implants induced the presence of disorganized scar tissue, while the Dual Mesh prostheses were encapsulated by organized tissue. The macrophage response was similar in both decreasing with time. The resistance to traction was higher when the reparation was done with polypropylene. We concluded that the structure of the prosthesis determines its degree of integration and the resistance to traction of the repaired zone.

One of the most serious problems with endothelial cell (EC) seeding of prosthetic materials is the poor adhesion and stability of the cells. Although several substrates that improve the initial adhesion have been assayed, the EC are lost within a limited period of time. In this study we attempted to modify the hydrophobic conditions of expanded polytetrafluoroethylene (ePTFE) by treating it with ethanol prior to seeding. In addition, we created a fibroblastic matrix that was also fixed by ethanol to the prosthetic material. In vitro studies were carried out at intervals of 24 hours and 15 days after seeding. EC from umbilical cord vein and fibroblasts from skin were seeded onto disks of PTFE with a porosity of 30 µm. The results obtained show that treatment of ePTFE with ethanol prior to EC seeding modifies its permeability, preventing cellular adhesion. The seeding of fibroblasts onto ePTFE allows a coating to form at 24 hours. The EC seeded onto this matrix adhere to it, forming a monolayer that persisted throughout the entire study period. The fibroblastic matrix allows the long-term survival of the EC on ePTFE.

Comparisons were made of the tissue response to the implantation of two different polytetrafluoroethylene prostheses: Soft Tissue Patch (STP) and Mycro Mesh (MM). A 7 × 5 cm prosthesis of STP (n=12) or MM (n=12) was implanted into a defect of the same size (involving all layers except skin) created in the anterior abdominal wall in 24 New Zealand rabbits. The prostheses were anchored to the recipient tissue, in direct contact with the intestinal loops and connective tissue. After 14, 30, 60 and 90 days, groups of six implants were studied macroscopically and samples were taken to be processed by light microscopy, scanning electron microscopy (SEM), immunohistochemical studies and tensiometry. All animals were valid for the study. In three cases STP implants presented very loose adhesions in the peripheral zones corresponding to the sutures. They were also observed on three MM implants, in the area of the perforations. Light and scanning microscopy revealed the formation of a capsule of scar tissue surrounding both types of prosthesis. At day 90, bridges of connective tissue had formed in the perforated areas of MM. Good vascularization was established in the areas of recipient tissue corresponding to both implants. The macrophage reaction to both biomaterials was maximal at 14 days, after which it progressively decreased until day 90. Tensile testing revealed no significant differences between the two biomaterials. It is concluded that (a) behaviour in the peritoneal interface is similar in the two prostheses, (b) both biomaterials become encapsulated rather than integrated into the recipient tissue, (c) the foreign body reaction does not determine the success or failure of the implants, (d) The perforations of the MM prosthesis do not increase its resistance to stress, or at least not after 90 days of implantation into rabbit abdominal wall.